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. Author manuscript; available in PMC: 2018 Dec 1.
Published in final edited form as: J Immunol. 2017 Oct 20;199(11):3757–3770. doi: 10.4049/jimmunol.1700822

Figure 2. The combination of BAFFR-Fc and anti-CD20 synergize in depleting B lymphocytes in the spleen and PLNs.

Figure 2

A) Frequency and absolute numbers of B220+ CD19+ B-lymphocytes amongst viable leukocytes in the spleens (left panel) and PLNs (middle panel) of NOD mice treated with ctrl mAb, BAFFR-Fc, anti-CD20, or combo from 10 to 14 weeks of age. Right panel depicts proportions of B-lymphocytes among CD45+ islet associated leukocytes from NOD mice treated with ctrl mAb, BAFFR-Fc, anti-CD20, or combo from 10 to 14 weeks of age. The total number of islets that could be isolated from each mouse varied. Thus, islet associated B-lymphocytes could only be assessed on a proportional basis. Data pooled from two independent experiments. B) Representative flow cytometric plots showing splenic T1 (CD21− CD23−), T2 (CD21hi CD23+), FO (CD21+ CD23+) and MZ (CD21hi CD23−) B-lymphocyte subsets among live CD19+ B220+ cells in NOD mice treated with ctrl mAb, BAFFR-Fc, anti-CD20, or combo regimens from 10 to 14 weeks of age (left panel). Absolute T1, T2, FO, MZ B-lymphocyte numbers calculated from values in the left panel (right panel). (n=10 per group). Data pooled from two independent experiments. C) Left panel-representative flow cytometric plots showing islet infiltrating B-lymphocytes co-expressing CD80 and PD-L2 among live CD19+ B220+ cells in NOD mice treated with ctrl mAb or BAFFR-Fc from 10 to 14 weeks of age. Right-panel-percent CD80+ PD-L2+ amongst splenic, PLN, or islet infiltrating B-lymphocytes in NOD mice treated with ctrl mAb or BAFFR-Fc from 10 to 14 weeks of age. (n=10/group for spleen/PLN); (n=5 mice/group for islets). Data pooled from two independent experiments for spleen and PLN. D) Primary or secondary HEL specific antibodies in NOD mice given ctrl mAb or BAFFR-Fc for 4 weeks starting at 10 weeks of age. (n=5 per group). p-values were calculated using Mann-Whitney analyses, each symbol represents an individual mouse; small horizontal lines indicate the Mean±SEM. Asterisks on top of each B-lymphocyte depleted groups indicate comparison to ctrl mAb (A-C). All other comparisons are drawn as lines between the indicated two groups. (**p<0.01, ***p<0.001, ****p<0.0001).